No abstract

Since 2008, the World Health Organization has provided seed grants to 11 manufacturers in low- and middle-income countries to establish or improve their pandemic influenza vaccine production capacity. To facilitate this ambitious project, an influenza vaccine technology platform (or "hub") was established at the Netherlands Vaccine Institute for training and technology transfer to developing countries. During its first two years of operation, a robust and transferable monovalent pilot process for egg-based inactivated whole virus influenza A vaccine production was established under international Good Manufacturing Practice standards, as well as in-process and release assays. A course curriculum was designed, including a two-volume practical handbook on production and quality control. Four generic hands-on training courses were successfully realized for over 40 employees from 15 developing country manufacturers. Planned extensions to the curriculum include cell-culture based technology for viral vaccine production, split virion influenza production, and generic adjuvant formulation. We conclude that technology transfer through the hub model works well, significantly builds vaccine manufacturing capacity in developing countries, and thereby increases global and equitable access to vaccines of high public health relevance.

Tissue-resident macrophages of the brain, including microglia, are implicated in the pathogenesis of various CNS disorders and are possible therapeutic targets by their chemical depletion or replenishment by hematopoietic stem cell therapy. Nevertheless, a comprehensive understanding of microglial function and the consequences of microglial depletion in the human brain is lacking. In human disease, heterozygous variants in CSF1R, encoding the Colony-stimulating factor 1 receptor, can lead to adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) possibly caused by microglial depletion. Here, we investigate the effects of ALSP-causing CSF1R variants on microglia and explore the consequences of microglial depletion in the brain. In intermediate- and late-stage ALSP post-mortem brain, we establish that there is an overall loss of homeostatic microglia and that this is predominantly seen in the white matter. By introducing ALSP-causing missense variants into the zebrafish genomic csf1ra locus, we show that these variants act dominant negatively on the number of microglia in vertebrate brain development. Transcriptomics and proteomics on relatively spared ALSP brain tissue validated a downregulation of microglia-associated genes and revealed elevated astrocytic proteins, possibly suggesting involvement of astrocytes in early pathogenesis. Indeed, neuropathological analysis and in vivo imaging of csf1r zebrafish models showed an astrocytic phenotype associated with enhanced, possibly compensatory, endocytosis. Together, our findings indicate that microglial depletion in zebrafish and human disease, likely as a consequence of dominant-acting pathogenic CSF1R variants, correlates with altered astrocytes. These findings underscore the unique opportunity CSF1R variants provide to gain insight into the roles of microglia in the human brain, and the need to further investigate how microglia, astrocytes, and their interactions contribute to white matter homeostasis.

Purpose: Recently, the Blue Mountains Eye Study reported an association between the use of topical timolol and cardiovascular mortality (Lee et al. Ophthalmology 2006). The purpose of the present study was to confirm or falsify this clinically very important finding, using data from the population‐based Rotterdam Study. Methods: 6971 participants of the Rotterdam Study, a longitudinal population based study of all residents aged 55 years and older from a district of Rotterdam, The Netherlands, were followed from 1991 onwards. Medication use and morbidity were recorded continuously during follow‐up. For the current analysis, baseline use of topical beta‐blockers and systemic cardiovascular medication as well as baseline cardiovascular morbidity were used, aiming to follow the design of the Blue Mountains Eye Study as close as possible. Cause of death was registered up to 1‐1‐2005. Data were analysed using Cox regression; Hazard ratios of topical beta‐blocker use were adjusted for age, sex, cardiovascular morbidity and use of systemic cardiovascular medication. Results: Mean age at baseline was 69 years (SD 9 years); 146 participants were using topical beta‐blockers at baseline. 2726 participants died during follow‐up (all cause mortality 40.1%), 611 (9.0%) had a cardiovascular cause of death. Hazard ratio of topical beta‐blocker use was 0.80 (95% confidence interval 0.63‐1.02; P=0.07) for all cause mortality and 0.78 (0.46‐1.29; P=0.32) for cardiovascular mortality. Conclusions: In our data, the use of topical beta‐blockers at baseline was not associated with either all cause mortality or cardiovascular mortality during follow‐up.